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Crossing strain varieties via swab to spore solution


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#41 DickMoby

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Posted 16 November 2020 - 08:54 AM

Hey sorry for putting out that energy.

 

Wasn't sure if I should write this comment because alot of people will read this, but felt like putting it here.

 

I tend to be very open and talk about things and tell my thoughts about them, often it gets ignored or results in uncomfortable situations, my intention tho is not to make anyone feel embarrassed or to police what people say (Someone told me I couldn't police what people say, on another platform).

__

 

I think to talk about facts at some points is important to prevent wrong info to be shared - this part is not about this thread here!

_________

 

Cultivation is what keeps me alive, the exchanges with other people for me is important part of it, often tho the situations I get into make me doubt myself and are hard to get out of.

Sometimes due to the speaking barrier there might be misinterpretations involved on both sides which makes it even harder.

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To add something to the actual thread.

 

I'm certain that I'm not the first one with this idea but what keeps people from cloning spore dropping PE fruits over some generations to get a fully spore dropping culture ?


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#42 fahtster

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Posted 16 November 2020 - 09:54 AM

You’re all good.. even if I don’t agree with all of it, I still value your input.. I think we could all use a little more perspective.

As for the PE.. I think workman said a long time ago that most PE drop spores.. it’s just usually really light and the you have to catch it at exactly the right time.. let me see if I can find it to quote.

Workman said:

“Here is the method which works most of the time. First, wait until the 3rd flush or later. Pick a nice looking mushroom that has dark gills. Timing is critical, if you see spores on the stem its probably too late for a good print. It takes a bit of luck or precognition to pick at the right time.

This is the secret part, shhhhhhh: twist and pull the stem out of the cap instead of slicing it off. This takes a bit of practice since the stem is tough and rough handling will crush the cap. If you cut off the stem instead of popping it out, the stump will block good spore fall. The stem has an odd swelling near the cap-stem junction. Print as you would any normal form of cubensis. Expect about a 20% success rate at getting relatively good prints. I'll post an example of what is considered a good PE print in a day or so.“

But that was also 14 years ago lol.. I recently saw someone get a really dark PE print on another site.. it just has to be at the right time and the window is really short.

It would be a nice project to do though, albeit a tough one with a lot of frustration I’m guessing

Faht
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#43 Freaky

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Posted 16 November 2020 - 12:25 PM

When I was printing PE I found it best to wait to full maturation (even a couple days after) though I didn't wait for later flushes.  Just keep an eye on the caps.  A good tell for PE is that they veil breaks early and ends up halfway down the stem once they're fully matured and that is a good indicator for me to watch for.  I would also take a handheld magnifier to look for light spore deposits so I could grab the caps with good deposit potential. 

 

I'm sure swabbing them is the best way to grab them now and most likely what I'd try if I wanted to collect spores from PE.


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#44 DickMoby

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Posted 16 November 2020 - 01:08 PM

The idea to spin the stem off is good.

 

What I plan to do tho is to only go for the fruits that drop good prints from alone.

To find them I'll just clone the ones that visibly have spores on their stems over as many generations as necessary to get a solid spore producing culture and then hopefully be able to take prints without any issues.

 

Maybe they print better with a little help like a glass over them for the printing or a drop of water on the cap like some people do with pans.

 

The magnifier sounds good aswell for the beginning of this process.

I could imagine that the idea of waiting for the later flushes is to print at the very end of the cultures life cycle when it puts the last bit of energy into reproduction (spore production).

 

I'm seriously interested to work that out to eliminate those annoying extra steps and to get a culture that at least has a high rate of naturally dropping spores, swabs are good as backup and easily taken but to send them in a normal letter to people for me at least, sending about half of the letters around half the globe is too risky to lose those letters (because you can feel the swabs through it).

 

Went again a little off topic here.

 

Btw if you disagree with something coming from me then feel free to talk about it.

I'm not here to put my opinion out and force everyone to take it, if you feel like your experience told you something different then I'm open to hear the other side to learn what went different with different subs or whatever it may be.

Also when talking generally about mush stuff, I'd call myself a little educated about the cultivation but especially with the background info online nobody is safe a 100% from wrong info, so if you see something that you disagree with then I'm always open to read and learn.


Edited by DickMoby, 16 November 2020 - 05:48 PM.

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#45 fahtster

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Posted 17 November 2020 - 04:08 PM

Those are jelly jars, but 1/2 pints should work fine too.. those jelly jars look full but there’s room in there for shaking. The reason I use the jelly jars is because the more grain you use, the water is going to get stuck in the jar because it sticks to the grain.. when I inject 20ml of water into the jelly jar, I get about 12ml back (That’s when the grain is hydrated—you’ll get more from a jar that has dried out grain—don’t let the water sit in the dried out grain super long either. It’ll absorb it pretty quickly)... 1/2 pints should be ok though.. just keep those things in mind.
Faht

 
Excellent! Thanks again Faht! And if I struggle with the half pints and issues I'll swap over to the smaller jelly jars.

I forgot to tell you that you should store the dried grains at room temp.. don’t put them in the fridge.. I made that mistake.. the glue from the tape will stay wet and the glue will mold.. I lost most of the jars I put in the fridge because if it.. i plan on trying to vacuum seal a jelly jar to see how well that works for extended periods (year+) but room temp In a drawer has always treated me right.

Faht
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#46 Freaky

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Posted 17 November 2020 - 06:23 PM

Thanks Faht, I definitely would of stored in a fridge and not swapped the lid out. 



#47 fahtster

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Posted 18 November 2020 - 10:01 AM

I also make my qt lids with the triple tyvek layer and over/under self-healing ports.76ED5918-F9A7-42C8-AE62-5EB358A6E755.jpeg

Run them ~ 10 times before I make new ones.. not because they fail but it’s nice to have new shiny things lol

Faht
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#48 Freaky

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Posted 18 November 2020 - 12:38 PM

I made up six - I tried my best to outline the inside but with those 3 layers I couldn't see through much.  Thanks for the tip on phone light - that should be bright enough to help.

 

Also I used electrical tape.  Made it really easy to wrap the band and the edges of the trimmed tyvek.  

 

Would it be better to run the silicone port along the center strip? I'm thinking like invert what I did - lay the tape on both sides and leave center strip open....I'm guessing any way I'd want to design the layout for tape and silicone is possible but to make sure that the tops and bottoms align for the silicone port.

 

Faht Lid1.jpg Faht Lid2.jpg

 

I do like them and think they'll work as well for me as they have for you.


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#49 fahtster

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Posted 18 November 2020 - 01:27 PM

Yeah.. that’s the best part.. you can make the design however you like.. just so you leave enough open for them to dry out.. I’m sure they’ll do great however you choose.

You can also use the fingers that you use to hold the lid while you Mark it to denote where the tape ends. Those look great! You can also use smaller silicone ports.. all up to you.. I just like the needle holes being far away from each other

Faht

Edited by fahtster, 18 November 2020 - 01:32 PM.

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#50 fahtster

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Posted 19 November 2020 - 11:38 AM

Pea+ Mini mono second flush.. these things are like sticks and matured very slowly, especially for second flushers (really fast fruits never fair well for potency ime) and they’re so pretty with that white bottom half of the caps

B3B09D83-3691-40A6-A7CD-5E0863455EFB.jpeg

Inoculated 4, f2 cakes yesterday.. used 5ml per cake to try and hurry the colonization along.. can’t wait to get these out for everyone

Faht
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#51 Freaky

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Posted 20 November 2020 - 06:48 PM

They are very pretty! I am excited to see the f2 version!

 

The cap coloring and damn they drop some dark spores - look at those veils are like a black tie - almost a tuxedo shroom you got going.


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#52 thafunkyone

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Posted 21 November 2020 - 05:57 AM

So, I'm trying to wrap my head around this- how exactly does putting 2 different mushrooms spores on the same swab make it a cross? you're basically just running a new MS grow, which would give you a genetic crapshoot since you're not isolating anything, but how does the genetic material blend? there is not sex involved, so there are no real 'parents' so to speak. everything we grow now is what it is through mycelial selection and cloning. wouldn't this just be starting with myc from whichever one won the genetic race (which we won't know)? all the PE variants, any albinos, were all selected, and growing conditions may change things a bit and cause lack of pigment or different colored spores etc, but there isn't any real 'breeding' or 'crossing' going on, just selecting the genetic traits that you're looking for and cloning.

 

I have been wrong before, I'm more wrong than right mostly, but this is the information I've been given over the last 20 years or so. You've been here longer than I have faht, and I'm sure you know more than me, so if I'm missing something here shoot me some links. I've been away for a couple years now so I have probably missed more than a little, but I was looking over the OMC and there are 'breeders' for mushrooms? how is this breeding? you can select and refine your selections to find something that fruits how you'd like and has beautiful rhizo growth etc, but mushrooms are nonsexual, they'll adapt to different substrates and growing conditions if you choose the tissue that prefers the environment you give it, but you cannot change the genetic makeup of something, especially through the genetic crapshoot known as multispore innoculation.



#53 DickMoby

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Posted 21 November 2020 - 09:07 AM

Because of exactly that for this method we HAVE TO choose two varieties with their own obvious characteristics.

 

The cross here has both genetics mixed up and therefore definately is a cross.

The spore production of AA+ with the mixed caps from PE.

A leucistic mushroom does not randomly turn into mixed caps mushrooms.

 

For PE the caps are also too different than usual and the spore production aswell.

 

Since the cross has traits mixed up that would normally not occur like that together, its definately a cross of both varieties.

 

Pasty did Rusty Whyte with swabs on agar, its the same principle.

You make sure that spores of both are so close together that (if they are compatible) some or maybe even just one strains is "created" by mating monos of both varieties.

 

Then by growing them out and cloning obvious crosses, the cross culture is selected and cloned over a few generations to get rid of other strains and then stabilized via printing of the crosses with the traits that you feel like are the ones you want.

 

With standard cubes that wouldn't be so easy to do, thats why I mentioned my mixed feelings about posting this tek - because maybe people do that with standard cubes and spread something that maybe isn't even a cross.

 

Here tho its obviously a mixture of both varieties


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#54 thafunkyone

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Posted 21 November 2020 - 01:21 PM

I get the theory, I'd like to see some actual science to back it up though, hence me asking for links. 

 

What about this theory, since I don't see hard science yet? 

 

My thoughts on this- all this stuff people call 'breeding' is selection, mostly of mutations, all the pigmentless, albino, different colored spored mushrooms  that have popped up are mutations. people would get random albino mushrooms in flushes and clone those suckers and fruit successive generations until that recessive or mutant trait became dominant. Now normal mushroom traits are rare in those selections. 

 

Now, lets throw some diversity in the mix by adding foreign spores. The confusion of all these spores germinating and competing makes it hard for the myc to keep all these selected traits that we've kept for so long and acts as a reset. Now we're back at square one. I don't see much in the way of PE in those, TBH. 

 

Thats my conclusion. If someone has data to show me otherwise I'd love it. I wouldn't call the mushrooms pictured above either name, because it doesn't resemble either of the originals. I don't think that's a bad thing though, if it does act as a reset it may have other recessive traitts in one of the 2 we don't know about and with selection we could have another unique mushroom, but I wouldn't call it a cross, unless someone can produce real actual science to back it up. 

 

The crossing question has come up a million times since I started doing this. People want new crosses just like weed, but flowers are sexual, mushrooms are asexual. I think that jumbling all these spores together get rid of all the recessive traits that we've tried to pull out of  the mushroooms and allowed one of the other sets of genetic instructions to control growth. 

 

I'm probably in the minority for these thoughts, but I go by what I've read and what the people that know more than me have told me. http://archives.myco...html?1075070998  is my reference link- page 333 deals with the topic at hand. It does discuss the crossing of different strains of the same mushroom and the fact that it can happen but generally will not produce fruitbodies, this is plainly producing fruits.

 

I'm not here to argue or call people stupid or anything, I just don't want misinformation to run rampant. Im' here with an open mind, if anyone has any links to back this up please send them my way, but until then I'll stand by my theory.


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#55 fahtster

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Posted 21 November 2020 - 01:43 PM

Yeah.. all that ^^^^ I specifically chose PE and AA+ because of the growth patterns of each... PE takes ~3 weeks to pin, whereas AA+ pins in a pretty normal timeframe.. with cakes, that timeframe isnt so easy to see, so I also made a mini mono.. the mini started pinning in 11 days after being spawned

PE has a very unique physical feature of those exaggerated bumps on the stem near the cap when mature.. a lot of, if not all, varieties’ fruits have these bumps but it’s usually barely there

Just the fact that I got normal colored fruits Pinning in 11 days with physical traits of AA+ (bell shaped, obvious two-toned caps) leads me to believe there’s been a cross. I’m still not at 100%, BUT I’m sure enough to say that it’s happened with confidence... but I’m moving on with it and going to f2 with cakes.. there I should see variations of the parents.. I don’t need to see full PE fruits (since they take longer to fruit—don’t need to wait).. all I need are the pea+ fruits and AA+ to be certain. The obvious pea+ fruits will be cloned and swabbed for f3

I wasn’t expecting to get mostly crossed fruits when I did this.. I was totally expecting to see AA+ pins and the ones that weren’t white pins but coming up at the same time as AA+, those were going to be my targets to watch.. but all I got were normal fruits which made me think I screwed up somewhere and mixed up swabs when I labeled them... but I’m a meticulous SOB and when I made the double swabs in April, I had this project in mind for this fall and I was very excited about it and while anything is possible as far as me making a mistake goes, im very doubtful of it and then I saw the physical characteristics of the mature fruits and now I’m confident once again that a mistake had not been made and the method of swabbing the spores over each other and then rubbing the two swabs together in sterile water, further entangling and meshing the two varieties’ spores next to each other was just so effective that all myc growth from that SS was growth from mostly mixed spores that mated

If it comes out that a mistake was made and these indeed are not a cross, I’ll course correct and own it, but I’m going on based on the evidence.. to me it’s pretty clear-cut. I knew this wouldn’t be easy going into it, that’s why I chose the varieties that I did.

Faht

#56 CatsAndBats

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Posted 21 November 2020 - 01:44 PM

Well then why am I harvesting all of this cottonmouth venom for?!

 

 

 

 

223-18-Fist-Shaking-Tantrum-Gif.gif


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#57 thafunkyone

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Posted 21 November 2020 - 02:05 PM

Yeah.. all that ^^^^ I specifically chose PE and AA+ because of the growth patterns of each... PE takes ~3 weeks to pin, whereas AA+ pins in a pretty normal timeframe.. with cakes, that timeframe isnt so easy to see, so I also made a mini mono.. the mini started pinning in 11 days after being spawned

PE has a very unique physical feature of those exaggerated bumps on the stem near the cap when mature.. a lot of, if not all, varieties’ fruits have these bumps but it’s usually barely there

Just the fact that I got normal colored fruits Pinning in 11 days with physical traits of AA+ (bell shaped, obvious two-toned caps) leads me to believe there’s been a cross. I’m still not at 100%, BUT I’m sure enough to say that it’s happened with confidence... but I’m moving on with it and going to f2 with cakes.. there I should see variations of the parents.. I don’t need to see full PE fruits (since they take longer to fruit—don’t need to wait).. all I need are the pea+ fruits and AA+ to be certain. The obvious pea+ fruits will be cloned and swabbed for f3

I wasn’t expecting to get mostly crossed fruits when I did this.. I was totally expecting to see AA+ pins and the ones that weren’t white pins but coming up at the same time as AA+, those were going to be my targets to watch.. but all I got were normal fruits which made me think I screwed up somewhere and mixed up swabs when I labeled them... but I’m a meticulous SOB and when I made the double swabs in April, I had this project in mind for this fall and I was very excited about it and while anything is possible as far as me making a mistake goes, im very doubtful of it and then I saw the physical characteristics of the mature fruits and now I’m confident once again that a mistake had not been made and the method of swabbing the spores over each other and then rubbing the two swabs together in sterile water, further entangling and meshing the two varieties’ spores next to each other was just so effective that all myc growth from that SS was growth from mostly mixed spores that mated

If it comes out that a mistake was made and these indeed are not a cross, I’ll course correct and own it, but I’m going on based on the evidence.. to me it’s pretty clear-cut. I knew this wouldn’t be easy going into it, that’s why I chose the varieties that I did.

Faht

 

I'm not here to attack- I'm here with an open mind. Let's see what the F2 brings! 

 

better send a print my way if they start getting wierd...:)


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#58 DickMoby

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Posted 21 November 2020 - 02:08 PM

Radical Mycology:

 

"RELATIONSHIP STRUCTURE AND SEXUAL ORIENTATION IN MUSHROOMS

While the pairing of two, sexually compatible partners is the most common relationship structure, it is not the rule. Not all Basidiomycete relationships are monogamous or capable of producing offspring. Occasionally, a partnered mycelium will invite in another mycelium, forming a triad. Two pairs of hyphae can also fuse to produce four-partner relationships, with each nucleus contributing its genetic knowledge and abilities to the healthy functioning of the system. This fungal version of polyamory, known as a polykaryon, can go on to create strong relationships and healthy offspring. Sometimes, a fusion occurs between partners who are somatically compatible, but infertile.
This is analogous to homosexuality in humans. These dikaryons can live together indefinitely, leading healthy lives. Without expending huge amounts of energy on reproduction, they have all the more life force to channel into their work of decomposition, symbiosis, or parasitis"
 
Not sure what you mean with asexual.
 
We got quite a few PE crosses out there:
PE6, Melmac, Tidal Wave, Kape, APE
 
those are the ones I know.
 
For example APE, its a cross of PF Albino and PE, both mutants.
Going with your theory APE could't exist how it exists today because both mutations would've "eliminated" each other.
 
If we take a look at all these crosses, they all don't have the classic PE dick look, something I see in the first pictures of this thread is wavy caps.
Some Melmacs and Tidal Wave (both PE crosses) do also have the wavy caps.
 
How would you explain the mixed fruits if not a cross?


#59 fahtster

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Posted 21 November 2020 - 02:13 PM

I keep getting nice little side effects from going from swab to SS.. so I made the f2 syringes on the 14th and didn’t get around to making the cakes for them until a couple days ago.. because of all the tiny bits of swab material with spores attached that comes off the tips due to the rubbing and ends up in the syringe, the spores germinated while the syringe sat.. the “tiny” bits turned into large bits when I went to inoculate the cakes.. the swab material is cotton and I used tap water so this makes sense. I already have myc growing after two days at all inoc points

B8A1EF70-72AC-4BAC-AB27-DFA7E1060903.jpeg

While this exciting, it also means that any contamination could have also fed off the cotton while it sat so I’m hoping the prewash did it’s job and that there wasn’t any mold spores mixed in with the mush spores that I grabbed with the swab. The prewash isn’t going to do mush for those. So fingers crossed!

Faht

#60 fahtster

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Posted 21 November 2020 - 02:17 PM

Yeah.. all that ^^^^ I specifically chose PE and AA+ because of the growth patterns of each... PE takes ~3 weeks to pin, whereas AA+ pins in a pretty normal timeframe.. with cakes, that timeframe isnt so easy to see, so I also made a mini mono.. the mini started pinning in 11 days after being spawned
PE has a very unique physical feature of those exaggerated bumps on the stem near the cap when mature.. a lot of, if not all, varieties’ fruits have these bumps but it’s usually barely there
Just the fact that I got normal colored fruits Pinning in 11 days with physical traits of AA+ (bell shaped, obvious two-toned caps) leads me to believe there’s been a cross. I’m still not at 100%, BUT I’m sure enough to say that it’s happened with confidence... but I’m moving on with it and going to f2 with cakes.. there I should see variations of the parents.. I don’t need to see full PE fruits (since they take longer to fruit—don’t need to wait).. all I need are the pea+ fruits and AA+ to be certain. The obvious pea+ fruits will be cloned and swabbed for f3
I wasn’t expecting to get mostly crossed fruits when I did this.. I was totally expecting to see AA+ pins and the ones that weren’t white pins but coming up at the same time as AA+, those were going to be my targets to watch.. but all I got were normal fruits which made me think I screwed up somewhere and mixed up swabs when I labeled them... but I’m a meticulous SOB and when I made the double swabs in April, I had this project in mind for this fall and I was very excited about it and while anything is possible as far as me making a mistake goes, im very doubtful of it and then I saw the physical characteristics of the mature fruits and now I’m confident once again that a mistake had not been made and the method of swabbing the spores over each other and then rubbing the two swabs together in sterile water, further entangling and meshing the two varieties’ spores next to each other was just so effective that all myc growth from that SS was growth from mostly mixed spores that mated
If it comes out that a mistake was made and these indeed are not a cross, I’ll course correct and own it, but I’m going on based on the evidence.. to me it’s pretty clear-cut. I knew this wouldn’t be easy going into it, that’s why I chose the varieties that I did.
Faht


I'm not here to attack- I'm here with an open mind. Let's see what the F2 brings!

better send a print my way if they start getting wierd...:)
I didn’t take it as an attack.. y’all will know if I’m upset lol trust me.. I’m all about discussion.. I was just explaining myself further.

Faht

Edited by fahtster, 21 November 2020 - 02:31 PM.





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